1. Field of the Invention
The present invention relates to Mycoplasma pneumoniae exotoxins, antibodies thereto, and their use in diagnostic and therapeutic methods.
2. Background Art
Mycoplasma pneumoniae is one of the most well recognized pathogens of the human respiratory tract. The importance of Mycoplasma pneumoniae cause of human respiratory disease has been well documented by epidemiological studies in various settings and in many countries. M. pneumoniae is the etiologic agent of primary atypical pneumonia and is also responsible for many respiratory tract infections, such as tracheobronchitis, bronchiolitis, pharyngitis and croup, especially in older children and young adults and in elderly populations. It accounts for 20-30% of all pneumonias and also is linked to asthma and chronic obstructive pulmonary disease. Furthermore, M. pneumoniae can disseminate to other organ sites and cause gastrointestinal, hematologic, neurologic, dermatologic, musculoskeletal and cardiovascular pathologies. This secondary involvement by M. pneumoniae leads to a spectrum of complicated extrapulmonary sequelae, including arthritis, pericarditis and central nervous system disorders, which attests to the significance of M. pneumoniae in human disease. Although antibiotic therapy appears to be relatively effective in controlling mycoplasma pneumonia, the bacteria continue to persist.
At present, no known virulence determinants of M. pneumoniae have been functionally identified and linked to the wide range of pathologies associated with M. pneumoniae mediated diseases. Furthermore, there are no specific and standardized diagnostic tests available for reliable and rapid detection of M. pneumoniae infection, or effective vaccines to control M pneumoniae infection.
The present invention overcomes previous shortcomings in the art by providing a Mycoplasma pneumoniae polypeptide and biologically active fragments thereof, known as community acquired respiratory distress syndrome (CARDS) toxin, as well as nucleic acids encoding this polypeptide and its fragments and antibodies specific thereto. These compositions are used, for example, in methods of diagnosing, treating and preventing infection by M. pneumoniae. 
Some sequences of this Invention:
Reference amino acid sequence M129/B9 (reference strain):(SEQ ID NO: 1)MPNPVRFVYR VDLRSPEEIF EHGFSTLGDV RNFFEHILST NFGRSYFIST SETPTAAIRF FGSWLREYVP EHPRRAYLYE IRADQHFYNA RATGENLLDL MRQRQVVFDS GDREMAQMGI RALRTSFAYQ REWFTDGPIA AANVRSAWLV DAVPVEPGHA HHPAGRVVET TRINEPEMHN PHYQELQTQA NDQPWLPTPG IATPVHLSIP QAASVADVSE GTSASLSFAC PDWSPPSSNG ENPLDKCIAE KIDNYNLQSL PQYASSVKEL EDTPVYLRGI KTQKTFMLQA DPQNNNVFLV EVNPKQKSSF PQTIFFWDVY QRICLKDLTG AQISLSLTAF TTQYAGQLKV HLSVSAVNAV NQKWKMTPQD IAITQFRVSS ELLGQTENGL FWNTKSGGSQ HDLYVCPLKN PPSDLEELQI IVDECTTHAQ FVTMRAASTF FVDVQLGWYW RGYYYTPQLS GWSYQMKTPD GQIFYDLKTS KIFFVQDNQN VFFLHNKLNK QTGYSWDWVE WLKHDMNEDK DENFKWYFSR DDLTIPSVEG LMFRHIRCYA DNQQLKVIIS GSRWGGWYST YDKVESNVED KILVKDGFDR F S1 (clinical strain) amino acid sequence: (SEQ ID NO: 2)MPNPVRFVYRVDLRSPEEIFEHGFSTLGDVRNFFEHIPSTNFGRSYFISTSETPTAAIRF FGSWLREYVPEHPRRAYLYEIRADQHFYNARATGENLLDLMRQRQVVFDSGDREMAQMGI RALRTSFAYQREWFTDGPIAAANVRSAWLVDAVPVEPGHAHHPAGRVVETTRINEPEMHN PHYQELQTQANDQPWLPTPGIATPVHLSIPQAASVADVSEGTSASLSFACPDWSPPSSNG ENPLDKCIAEKIDNYNLQSLPQYASSVKELEDTPVYLRGIKTQKTFMLQADPQNNNVFLV EVNPKQKSPFPQTIFFWDVYQRICLKDLTGAQISLSLTAFTTQYAGQLKVHLSVSAVNAV NQKWKMTPQDSAITQFRVSSELLGQTENGLSWNTKSGGSQHDLYVCPLKNPPSDLEELQI IVDECTTHAQFVTMRAASTFFVDVQLGWYWRGYYYTPQLSGWSYQMKTPDGQIFYDLKTS KIFFVQDNQNVFFLHNKLNKQTGYSWDWVEWLKHDMNEDKDENFKWYFSRDDLTIPSVEG LNFRHIRCYADNQQLKVIISGSRWGGWYSTYDKVESNVEDKILVKDGFDRF JL (clinical strain) amino acid sequence: (SEQ ID NO: 3)MPNPVRFVYRVDLRSPEEIFEHGFSTLGDVRNFFEHILSTNFGRSYFISTSETPTAAIRF FGSWLREYVPEHPRRAYLYEIRADQHFYNARATGENLLDLMRQRQVVFDSGDREMAQMGI RALRTSFAYQREWFTDGPIAAANVRSAWLVDAVPVEPGHAHHPAGRVVETTRINEPEMHN PHYQELQTQANDQPWLPTPGIATPVHLSIPQAASVADVSEGTSASLSFACPDWSPPSSNG ENPLDKCIAEKIDNYNLQSLPQYASSVKELEDTPVYLRGIKTQKTFMLQADPQNNNVFLV EVNPKQKSSFPQTIFFWDVYQRICLKDLTGAQISLSLTAFTTQYAGQLKVHLSVSAVNAV NQKWKMTPQDSAITQFRVSSELLGQTENGLFWNTKSGGSQHDLYVCPLKNPPSDLEELQI IVDECTTHAQFVTMRAASTFFVDVQLGWYWRGYYYTPQLSGWSYQMKTPDGQIFYDLKTS KIFFVQDNQNVFFLHNKLNKQTGYSWDWVEWLKHDMNEDKDENFKWYFSRDDLTIPSVEG LNFRHIRCYADNQQLKVIISGSRWGGWYSTYDKVESNVEDKILVKDGFDRF RJL1 (clinical strain) amino acid sequence: (SEQ ID NO: 4)MPNPVRFVYRVDLRSPEEIFEHGFSTLGDVRNFFEHILSTNFGRSYFISTSETPTAAIRF FGSWLREYVPEHPRRAYLYEIRADQHFYNARATGENLLDLMRQRQVVFDSGDREMAQMGI RALRTSFAYQREWFTDGPIAAANVRSAWLVDAVPVEPGHAHHPAGRVVETTRINEPEMHN PHYQELQTQANDQPWLPTPGIATPVHLSIPQAASVADVSEGTSASLSFACPDWSPPSSNG ENPLDKCIAEKIDNYNLQSLPQYASSVKELEDTPVYLRGIKTQKTFMLQADPQNNNVFLV EVNPKQKSSFPQTIFFWDVYQRICLKDLTGAQISLSLTAFTTQYAGQLKVHLSVSAVNAV NQKWKMTPQDSAITQFRVSSELLGQTENGLFRNTKSGGSQHDLYVCPLKNPPSDLEELQI IVDECTTHAQFVTMRAASTFFVDVQLGWYWRGYYYTPQLSGWSYQMKTPDGQIFYDLKTS KIFFVQDNQNVFFLHNKLNKQTGYSWDWVEWLKHDMNEDKDENFKWYFSRDDLTIPSVEG LNFRHIRCYADNQQLKVIISGSRWGGWYSTYDKVESNVEDKILVKDGFDRF L2 (clinical strain) amino acid sequence: (SEQ ID NO: 5)MPNPVRFVYRVDLRSPEEIFEHGFSTLGDVRNFFEHILSTNFGRSYFISTSETPTAAIRF FGSWLREYVPEHPRRAYLYEIRADQHFYNARATGENLLDLMRQRQVVFDSGDREMAQMGI RALRTSFAYQREWFTDGPIAAANVRSAWLVDAVPVEPGHAHHPAGRVVETTRINEPEMHN PHYQELQTQANDQPWLPTPGIATPVHLSIPQAASVADVSEGTSASLSFACPDWSPPSSNG ENPLGKCIAEKIDNYNLQSLPQYASSVKELEDTPVYLRGIKTQKTFMLQADPQNNNVFLV EVNPKQKSSFPQTIFFWDVYQRICLKDLTGAQISLSLTAFTTQYAGQLKVHLSVSAVNAV NQKWKMTPQDSAITQFRVSSELLGQTENGLFWNTKSGGSQHDLYVCPLKNPPSDLEELQI IVDECTTHAQFVTMRAASTFFVDVQLGWYWRGYYYTPQLSGWSYQMKTPDGQIFYDLKTS KIFFVQDNQNVFFLHNKLNKQTGYSWDWVEWLKHDMNEDKDENFKWYFSRDDLTIPSVEG LNFRHIRCYADNQQLKVIISGSRWGGWYSTYDKVESNVEDKILVKDGFDRF Composite amino acid sequence: (SEQ ID NO: 6)MPNPVRFVYR VDLRSPEEIF EHGFSTLGDV RNFFEHIPST NFGRSYFIST SETPTAAIRF FGSWLREYVP EHPRRAYLYE IRADQHFYNA RATGENLLDL MRQRQVVFDS GDREMAQMGI RALRTSFAYQ REWFTDGPIA AANVRSAWLV DAVPVEPGHA HHPAGRVVET TRINEPEMHN PHYQELQTQA NDQPWLPTPG IATPVHLSIP QAASVADVSE GTSASLSFAC PDWSPPSSNG ENPLGKCIAE KIDNYNLQSL PQYASSVKEL EDTPVYLRGI KTQKTFMLQA DPQNNNVFLV EVNPKQKPSF PQTIFFWDVY QRICLKDLTG AQISLSLTAF TTQYAGQLKV HLSVSAVNAV NQKWKMTPQD SAITQFRVSS ELLGQTENGL SRNTKSGGSQ HDLYVCPLKN PPSDLEELQI IVDECTTHAQ FVTMRAASTF FVDVQLGWYW RGYYYTPQLS GWSYQMKTPD GQIFYDLKTS KIFFVQDNQN VFFLHNKLNK QTGYSWDWVE WLKHDMNEDK DENFKWYFSR DDLTIPSVEG LNFRHIRCYA DNQQLKVIIS GSRWGGWYST YDKVESNVED KILVKDGFDR F Reference nucleotide sequence M129/B9 (contains tga's that needto be changed to tgg before expression in E. coli)(SEQ ID NO: 7)tttttaattt gtaaaatttc attttttaaa aatgccaaat cctgttagat ttgtttaccg tgttgatttg agaagccctg aagaaatttt tgaacatggc ttttcaactt taggtgatgt gagaaatttc tttgaacaca ttctctccac taattttggt agaagctatt ttatttccac ttcagaaaca cccacagcag ctattcgctt ctttggtagc tggttacggg aatatgtacc agagcacccc agaagggctt acttatatga aattcgtgcc gaccaacact tttacaatgc ccgcgccact ggggagaact tgttagattt aatgcgtcaa agacaagtag tatttgactc tggtgatcga gaaatggcac aaatgggaat tagagcttta cgcacttcct ttgcgtatca acgtgaatgg tttaccgatg gtccaattgc agcagctaat gtccgtagtg cttgactagt agatgctgtt cccgttgaac ctggtcatgc tcaccacccg gctggtcgtg ttgtagagac tactagaatt aatgaaccgg aaatgcacaa ccctcattat caagagctgc aaacccaagc caatgatcaa ccatgattgc caacaccagg aatagctact cctgtacatt tatcaattcc ccaagcagct tccgttgctg atgtttcgga aggtacttcc gcttcgctat cgtttgcgtg ccctgattga agtccacctt ctagtaatgg tgaaaatccg ctagacaaat gcattgcgga aaagattgat aactataacc tacaatcctt accacagtac gctagcagtg taaaggaact ggaagataca ccagtatacc taaggggaat taaaacgcaa aaaaccttta tgttacaagc agatccgcaa aataacaatg tctttttggt cgaagtaaac cccaaacaaa agtccagctt tccccaaacc atcttctttt gggatgttta tcaacgaatt tgtctcaagg atttaactgg tgcacaaatc agtctttcgc ttactgcctt tactactcag tatgctggtc agctcaaagt gcaccttagt gttagcgcgg ttaatgccgt gaaccaaaag tgaaaaatga caccgcaaga cattgcaata actcagtttc gggtctcctc tgaactgtta ggtcaaactg aaaatggctt gttctgaaat accaagagtg gtggttcaca acacgatttg tatgtatgtc ctttgaaaaa tccacctagt gatttggaag aattacaaat aattgttgat gaatgtacta cccatgcgca gtttgttact atgcgtgcag ctagcacctt ctttgttgat gttcagctag gctggtattg aaggggttat tactataccc cacaattaag tggttgatct tatcagatga aaacaccaga tggacagata ttctatgatc taaaaacttc gaaaatcttc tttgtccagg acaaccaaaa cgtgttcttt ctccataata aactcaacaa acaaactggt tacagctggg attgagtaga atggctaaaa catgacatga atgaggacaa agacgaaaac tttaaatggt acttttcgcg tgatgacctt accattcctt ccgttgaagg gcttaacttc cgccacattc gctgttacgc tgacaaccag cagttaaagg tgatcataag cggttcacgt tggggcggtt ggtactccac ttacgataaa gttgaaagta atgtcgaaga taagattttg gtcaaagatg gttttgatcg cttttagcga ttaagcttta acgtcactgt tttgctctaa tgttagaagc aaagatcttg S1 Nucleotide sequence with each tga changed to tgg for expres-sion in E. coli (SEQ ID NO: 8)atgccaaatc ctgttagatt tgtttaccgt gttgatttga gaagccctga agaaattttt60 gaacatggct tttcaacttt aggtgatgtg agaaatttct ttgaacacat tccctccact120 aattttggta gaagctattt tatttccact tcagaaacac ccacagcagc tattcgcttc180 tttggtagct ggttacggga atatgtacca gagcacccca gaagggctta cttatatgaa240 attcgtgccg accaacactt ttacaatgcc cgcgccactg gggagaactt gttagattta300 atgcgtcaaa gacaagtagt atttgactct ggtgatcgag aaatggcaca aatgggaatt360 agagctttac gcacttcctt tgcgtatcaa cgtgaatggt ttaccgatgg tccaattgca420 gcagctaatg tccgtagtgc ttggctagta gatgctgttc ccgttgaacc tggtcatgct480 caccacccgg ctggtcgtgt tgtagagact actagaatta atgaaccgga aatgcacaac540 cctcattatc aagagctgca aacccaagcc aatgatcaac catggttgcc aacaccagga600 atagctactc ctgtacattt atcaattccc caagcagctt ccgttgctga tgtttcggaa660 ggtacttccg cttcgctatc gtttgcgtgc cctgattgga gtccaccttc tagtaatggt720 gaaaatccgc tagacaaatg cattgcggaa aagattgata actataacct acaatcctta780 ccacagtacg ctagcagtgt aaaggaactg gaagatacac cagtatacct aaggggaatt840 aaaacgcaaa aaacctttat gttacaagca gatccgcaaa ataacaatgt ctttttggtc900 gaagtaaacc ccaaacaaaa gcccagcttt ccccaaacca tcttcttttg ggatgtttat960 caacgaattt gtctcaagga tttaactggt gcacaaatca gtctttcgct tactgccttt1020 actactcagt atgctggtca gctcaaagtg caccttagtg ttagcgcggt taatgccgtg1080 aaccaaaagt ggaaaatgac accgcaagac agtgcaataa ctcagtttcg ggtctcctct1140 gaactgttag gtcaaactga aaatggcttg tcctggaata ccaagagtgg tggttcacaa1200 cacgatttgt atgtatgtcc tttgaaaaat ccacctagtg atttggaaga attacaaata1260 attgttgatg aatgtactac ccatgcgcag tttgttacta tgcgtgcagc tagcaccttc1320 tttgttgatg ttcagctagg ctggtattgg aggggttatt actatacccc acaattaagt1380 ggttggtctt atcagatgaa aacaccagat ggacagatat tctatgatct aaaaacttcg1440 aaaatcttct ttgtccagga caaccaaaac gtgttctttc tccataataa actcaacaaa1500 caaactggtt acagctggga ttgggtagaa tggctaaaac atgacatgaa tgaggacaaa1560 gacgaaaact ttaaatggta cttttcgcgt gatgacctta ccattccttc cgttgaaggg1620 cttaacttcc gccacattcg ctgttacgct gacaaccagc agttaaaggt gatcataagc1680 ggttcacgtt ggggcggttg gtactccact tacgataaag ttgaaagtaa tgtcgaagat1740 aagattttgg tcaaagatgg ttttgatcgc ttt1773 L2 nucleotide sequence with each tga changed to tgg for expression inE. coli (SEQ ID NO: 9)atgccaaatc ctgttagatt tgtttaccgt gttgatttga gaagccctga agaaattttt60 gaacatggct tttcaacttt aggtgatgtg agaaatttct ttgaacacat tctctccact120 aattttggta gaagctattt tatttccact tcagaaacac ccacagcagc tattcgcttc180 tttggtagct ggttacggga atatgtacca gagcacccca gaagggctta cttatatgaa240 attcgtgccg accaacactt ttacaatgcc cgcgccactg gggagaactt gttagattta300 atgcgtcaaa gacaagtagt atttgactct ggtgatcgag aaatggcaca aatgggaatt360 agagctttac gcacttcctt tgcgtatcaa cgtgaatggt ttaccgatgg tccaattgca420 gcagctaatg tccgtagtgc ttggctagta gatgctgttc ccgttgaacc tggtcatgct480 caccacccgg ctggtcgtgt tgtagagact actagaatta atgaaccgga aatgcacaac540 cctcattatc aagagctgca aacccaagcc aatgatcaac catggttgcc aacaccagga600 atagctactc ctgtacattt atcaattccc caagcagctt ccgttgctga tgtttcggaa660 ggtacttccg cttcgctatc gtttgcgtgc cctgattgga gtccaccttc tagtaatggt720 gaaaatccgc taggcaaatg cattgcggaa aagattgata actataacct acaatcctta780 ccacagtacg ctagcagtgt aaaggaactg gaagatacac cagtatacct aaggggaatt840 aaaacgcaaa aaacctttat gttacaagca gatccgcaaa ataacaatgt ctttttggtc900 gaagtaaacc ccaaacaaaa gtccagcttt ccccaaacca tcttcttttg ggatgtttat960 caacgaattt gtctcaagga tttaactggt gcacaaatca gtctttcgct tactgccttt1020 actactcagt atgctggtca gctcaaagtg caccttagtg ttagcgcggt taatgccgtg1080 aaccaaaagt ggaaaatgac accgcaagac agtgcaataa ctcagtttcg ggtctcctct1140 gaactgttag gtcaaactga aaatggcttg ttctggaata ccaagagtgg tggttcacaa1200 cacgatttgt atgtatgtcc tttgaaaaat ccacctagtg atttggaaga attacaaata1260 attgttgatg aatgtactac ccatgcgcag tttgttacta tgcgtgcagc tagcaccttc1320 tttgttgatg ttcagctagg ctggtattgg aggggttatt actatacccc acaattaagt1380 ggttggtctt atcagatgaa aacaccagat ggacagatat tctatgatct aaaaacttcg1440 aaaatcttct ttgtccagga caaccaaaac gtgttctttc tccataataa actcaacaaa1500 caaactggtt acagctggga ttgggtagaa tggctaaaac atgacatgaa tgaggacaaa1560 gacgaaaact ttaaatggta cttttcgcgt gatgacctta ccattccttc cgttgaaggg1620 cttaacttcc gccacattcg ctgttacgct gacaaccagc agttaaaggt gatcataagc1680 ggttcacgtt ggggcggttg gtactccact tacgataaag ttgaaagtaa tgtcgaagat1740 aagattttgg tcaaagatgg ttttgatcgc ttt1773 JL nucleotide sequence with each tga changed to tgg for expression inE. coli (SEQ ID NO: 10)atgccaaatc ctgttagatt tgtttaccgt gttgatttga gaagccctga agaaattttt60 gaacatggct tttcaacttt aggtgatgtg agaaatttct ttgaacacat tctctccact120 aattttggta gaagctattt tatttccact tcagaaacac ccacagcagc tattcgcttc180 tttggtagct ggttacggga atatgtacca gagcacccca gaagggctta cttatatgaa240 attcgtgccg accaacactt ttacaatgcc cgcgccactg gggagaactt gttagattta300 atgcgtcaaa gacaagtagt atttgactct ggtgatcgag aaatggcaca aatgggaatt360 agagctttac gcacttcctt tgcgtatcaa cgtgaatggt ttaccgatgg tccaattgca420 gcagctaatg tccgtagtgc ttggctagta gatgctgttc ccgttgaacc tggtcatgct480 caccacccgg ctggtcgtgt tgtagagact actagaatta atgaaccgga aatgcacaac540 cctcattatc aagagctgca aacccaagcc aatgatcaac catggttgcc aacaccagga600 atagctactc ctgtacattt atcaattccc caagcagctt ccgttgctga tgtttcggaa660 ggtacttccg cttcgctatc gtttgcgtgc cctgattgga gtccaccttc tagtaatggt720 gaaaatccgc tagacaaatg cattgcggaa aagattgata actataacct acaatcctta780 ccacagtacg ctagcagtgt aaaggaactg gaagatacac cagtatacct aaggggaatt840 aaaacgcaaa aaacctttat gttacaagca gatccgcaaa ataacaatgt ctttttggtc900 gaagtaaacc ccaaacaaaa gtccagcttt ccccaaacca tcttcttttg ggatgtttat960 caacgaattt gtctcaagga tttaactggt gcacaaatca gtctttcgct tactgccttt1020 actactcagt atgctggtca gctcaaagtg caccttagtg ttagcgcggt taatgccgtg1080 aaccaaaagt ggaaaatgac accgcaagac agtgcaataa ctcagtttcg ggtctcctct1140 gaactgttag gtcaaactga aaatggcttg ttctggaata ccaagagtgg tggttcacaa1200 cacgatttgt atgtatgtcc tttgaaaaat ccacctagtg atttggaaga attacaaata1260 attgttgatg aatgtactac ccatgcgcag tttgttacta tgcgtgcagc tagcaccttc1320 tttgttgatg ttcagctagg ctggtattgg aggggttatt actatacccc acaattaagt1380 ggttggtctt atcagatgaa aacaccagat ggacagatat tctatgatct aaaaacttcg1440 aaaatcttct ttgtccagga caaccaaaac gtgttctttc tccataataa actcaacaaa1500 caaactggtt acagctggga ttgggtagaa tggctaaaac atgacatgaa tgaggacaaa1560 gacgaaaact ttaaatggta cttttcgcgt gatgacctta ccattccttc cgttgaaggg1620 cttaacttcc gccacattcg ctgttacgct gacaaccagc agttaaaggt gatcataagc1680 ggttcacgtt ggggcggttg gtactccact tacgataaag ttgaaagtaa tgtcgaagat1740 aagattttgg tcaaagatgg ttttgatcgc ttt1773 RJL1 nucleotide sequence with each tga changed to tgg for expressionin E. coli (SEQ ID NO: 11)atgccaaatc ctgttagatt tgtttaccgt gttgatttga gaagccctga agaaattttt60 gaacatggct tttcaacttt aggtgatgtg agaaatttct ttgaacacat tctctccact120 aattttggta gaagctattt tatttccact tcagaaacac ccacagcagc tattcgcttc180 tttggtagct ggttacggga atatgtacca gagcacccca gaagggctta cttatatgaa240 attcgtgccg accaacactt ttacaatgcc cgcgccactg gggagaactt gttagattta300 atgcgtcaaa gacaagtagt atttgactct ggtgatcgag aaatggcaca aatgggaatt360 agagctttac gcacttcctt tgcgtatcaa cgtgaatggt ttaccgatgg tccaattgca420 gcagctaatg tccgtagtgc ttggctagta gatgctgttc ccgttgaacc tggtcatgct480 caccacccgg ctggtcgtgt tgtagagact actagaatta atgaaccgga aatgcacaac540 cctcattatc aagagctgca aacccaagcc aatgatcaac catggttgcc aacaccagga600 atagctactc ctgtacattt atcaattccc caagcagctt ccgttgctga tgtttcggaa660 ggtacttccg cttcgctatc gtttgcgtgc cctgattgga gtccaccttc tagtaatggt720 gaaaatccgc tagacaaatg cattgcggaa aagattgata actataacct acaatcctta780 ccacagtacg ctagcagtgt aaaggaactg gaagatacac cagtatacct aaggggaatt840 aaaacgcaaa aaacctttat gttacaagca gatccgcaaa ataacaatgt ctttttggtc900 gaagtaaacc ccaaacaaaa gtccagcttt ccccaaacca tcttcttttg ggatgtttat960 caacgaattt gtctcaagga tttaactggt gcacaaatca gtctttcgct tactgccttt1020 actactcagt atgctggtca gctcaaagtg caccttagtg ttagcgcggt taatgccgtg1080 aaccaaaagt ggaaaatgac accgcaagac agtgcaataa ctcagtttcg ggtctcctct1140 gaactgttag gtcaaactga aaatggcttg ttccggaata ccaagagtgg tggttcacaa1200 cacgatttgt atgtatgtcc tttgaaaaat ccacctagtg atttggaaga attacaaata1260 attgttgatg aatgtactac ccatgcgcag tttgttacta tgcgtgcagc tagcaccttc1320 tttgttgatg ttcagctagg ctggtattgg aggggttatt actatacccc acaattaagt1380 ggttggtctt atcagatgaa aacaccagat ggacagatat tctatgatct aaaaacttcg1440 aaaatcttct ttgtccagga caaccaaaac gtgttctttc tccataataa actcaacaaa1500 caaactggtt acagctggga ttgggtagaa tggctaaaac atgacatgaa tgaggacaaa1560 gacgaaaact ttaaatggta cttttcgcgt gatgacctta ccattccttc cgttgaaggg1620 cttaacttcc gccacattcg ctgttacgct gacaaccagc agttaaaggt gatcataagc1680 ggttcacgtt ggggcggttg gtactccact tacgataaag ttgaaagtaa tgtcgaagat1740 aagattttgg tcaaagatgg ttttgatcgc ttt1773